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Orbital shaker

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The Orbital shaker is a laboratory equipment designed to provide gentle agitation for a variety of applications. It features a stable platform that can accommodate various types of containers, allowing for consistent and controlled mixing of samples.

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Lab products found in correlation

Ultra low attachment 6 well plate, by Corning (4 mentions) Growth factor reduced matrigel, by Corning (4 mentions) 96 well u bottom plate, by Corning (2 mentions) Mem neaa, by Corning (2 mentions) Dmem f12, by Thermo Fisher Scientific (2 mentions) N2 supplement, by Thermo Fisher Scientific (2 mentions) Glutamax supplement, by Thermo Fisher Scientific (2 mentions) Mem neaa, by Thermo Fisher Scientific (2 mentions) Heparin, by Thermo Fisher Scientific (2 mentions) Insulin, by Thermo Fisher Scientific (2 mentions) Ultralow attachment u bottom 96 well plates, by Corning (2 mentions) Neurobasal, by Thermo Fisher Scientific (1 mentions)

6 protocols using orbital shaker

1

Cerebral Organoid Generation and Culture

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To generate COs, 1 × 104 single cell-dissociated hESCs were plated on each well of ultralow-attachment U-bottom 96-well plates (Corning). At 24 hr after plating, BGM supplemented with 2 μM dorsomorphin and 2 μM A83-01 were added to EBs. The medium was replaced every other day. After 7 days of differentiation, COs were embedded into growth factor-reduced matrigel (Corning) droplets. Matrigel-embedded COs were transferred to 6-cm petri dishes containing BGM supplemented with 200 ng/ml laminin and 2.5 μg/ml insulin for inducing basal-apical lamination. After 2 days, COs were transferred into ultralow-attachment 6-well plates (Corning) containing BMM and cultured on an orbital shaker (Stuart). The culture medium was replaced every other day.
Yao X., Kang J.H., Kim K.P., Shin H., Jin Z.L., Guo H., Xu Y.N., Li Y.H., Hali S., Kwon J., La H., Park C., Kim Y.J., Wang L., Hong K., Cao Q., Cho I.J., Kim N.H, & Han D.W. (2023). Production of Highly Uniform Midbrain Organoids from Human Pluripotent Stem Cells. Stem Cells International, 2023, 3320211.
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2

Generation of Brain Organoids from hESCs

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Brain organoids were generated from hESCs as previously described 39 (link), but slightly modified. Briefly, for embryoid body (EB) formation, hESCs were washed twice with DPBS, incubated with Accutase for 5 minutes, and dissociated into single cells. 3000 single cells were seeded in each well of low attachment 96-well U-bottom plate in E8 medium containing 10 μM ROCK inhibitor and centrifuged at 100 g for 3 min, then medium was half changed every other day. On day 4, EBs were transferred to low attachment 24-well plate in neural induction medium containing DMEM-F12 (GIBCO) with 1% N2 supplement (GIBCO), 1% Glutamax supplement (GIBCO), 1% MEM-NEAA (GIBCO) and 1 μg/mL Heparin (GIBCO), and medium was changed after 48 h. On day 7, EBs were transferred into Matrigel droplets as previously described and cultured in brain organoid differentiation media containing 50% DMEM-F12, 50% Neurobasal, 200x N2 supplement, 0.025% Insulin (GIBCO), 100x Glutamax supplement, 200x MEM-NEAA, 100x penicillin-streptomycin, 0.035% 2-Mercaptoethanol and 100x B27 supplement without Vitamin A, and medium was changed after 48 h. On day 10, organoids were transferred to orbital shaker (Corning) in brain organoid differentiation media with Vitamin A, medium was changed every 4 days.
Zhang X., Lin H., Dong L, & Xia Q. (2022). Recapitulating influenza virus infection and facilitating antiviral and neuroprotective screening in tractable brain organoids. Theranostics, 12(12), 5317-5329.
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3

Cerebral Organoid Generation and Culture

Check if the same lab product or an alternative is used in the 5 most similar protocols
To generate COs, 1 × 104 single cell-dissociated hESCs were plated on each well of ultralow-attachment U-bottom 96-well plates (Corning). At 24 hr after plating, BGM supplemented with 2 μM dorsomorphin and 2 μM A83-01 were added to EBs. The medium was replaced every other day. After 7 days of differentiation, COs were embedded into growth factor-reduced matrigel (Corning) droplets. Matrigel-embedded COs were transferred to 6-cm petri dishes containing BGM supplemented with 200 ng/ml laminin and 2.5 μg/ml insulin for inducing basal-apical lamination. After 2 days, COs were transferred into ultralow-attachment 6-well plates (Corning) containing BMM and cultured on an orbital shaker (Stuart). The culture medium was replaced every other day.
Yao X., Kang J.H., Kim K.P., Shin H., Jin Z.L., Guo H., Xu Y.N., Li Y.H., Hali S., Kwon J., La H., Park C., Kim Y.J., Wang L., Hong K., Cao Q., Cho I.J., Kim N.H, & Han D.W. (2023). Production of Highly Uniform Midbrain Organoids from Human Pluripotent Stem Cells. Stem Cells International, 2023, 3320211.
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4

Hindbrain Organoid Generation from hESCs

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To generate hindbrain organoids (HOs), 1 × 104 single cell-dissociated hESCs were plated on each well of ultralow-attachment U-bottom 96-well plates (Corning). At 24 hr after plating, BGM supplemented with 2 μM dorsomorphin, 2 μM A83-01, and 4.25 μM CHIR99021was added to EBs. The medium was replaced every other day. After 7 days of differentiation, HOs were embedded into growth factor-reduced matrigel (Corning) droplets. Matrigel-embedded HOs were transferred to 6-cm petri dishes. After 2 days, HOs were transferred into ultralow-attachment 6-well plates (Corning) containing BMM and cultured on an orbital shaker (Stuart). The culture medium was replaced every other day.
Yao X., Kang J.H., Kim K.P., Shin H., Jin Z.L., Guo H., Xu Y.N., Li Y.H., Hali S., Kwon J., La H., Park C., Kim Y.J., Wang L., Hong K., Cao Q., Cho I.J., Kim N.H, & Han D.W. (2023). Production of Highly Uniform Midbrain Organoids from Human Pluripotent Stem Cells. Stem Cells International, 2023, 3320211.
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5

Hindbrain Organoid Generation from hESCs

Check if the same lab product or an alternative is used in the 5 most similar protocols
To generate hindbrain organoids (HOs), 1 × 104 single cell-dissociated hESCs were plated on each well of ultralow-attachment U-bottom 96-well plates (Corning). At 24 hr after plating, BGM supplemented with 2 μM dorsomorphin, 2 μM A83-01, and 4.25 μM CHIR99021was added to EBs. The medium was replaced every other day. After 7 days of differentiation, HOs were embedded into growth factor-reduced matrigel (Corning) droplets. Matrigel-embedded HOs were transferred to 6-cm petri dishes. After 2 days, HOs were transferred into ultralow-attachment 6-well plates (Corning) containing BMM and cultured on an orbital shaker (Stuart). The culture medium was replaced every other day.
Yao X., Kang J.H., Kim K.P., Shin H., Jin Z.L., Guo H., Xu Y.N., Li Y.H., Hali S., Kwon J., La H., Park C., Kim Y.J., Wang L., Hong K., Cao Q., Cho I.J., Kim N.H, & Han D.W. (2023). Production of Highly Uniform Midbrain Organoids from Human Pluripotent Stem Cells. Stem Cells International, 2023, 3320211.
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6

Generating Brain Organoids from hESCs

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Brain organoids were generated from hESCs as previously described (Lancaster and Knoblich, 2014) , but slightly modified. Briefly, for embryoid body (EB) formation, hESCs were washed twice with DPBS, incubated with Accutase for 5 minutes, and dissociated into single cells. 3000 single cells were seeded in each well of low attachment 96-well U-bottom plate in E8 medium containing 10 µM ROCK inhibitor and centrifuged at 100 g for 3 min, then medium was half changed every other day. On day 4, EBs were transferred to low attachment 24-well plate in neural induction medium containing DMEM-F12 (GIBCO) with 1% N2 supplement (GIBCO), 1% Glutamax supplement (GIBCO), 1% MEM-NEAA (GIBCO) and 1 µg/mL Heparin (GIBCO), and medium was changed after 48 h. On day 7, EBs were transferred into Matrigel droplets as previously described and cultured in brain organoid differentiation media containing 50% DMEM-F12, 50%
Neurobasal, 200x N2 supplement, 0.025% Insulin (GIBCO), 100x Glutamax supplement, 200x MEM-NEAA, 100x penicillin-streptomycin, 0.035% 2-Mercaptoethanol and 100x B27 supplement without Vitamin A, and medium was changed after 48 h. On day 10, organoids were transferred to orbital shaker (Corning) in brain organoid differentiation media with Vitamin A, medium was changed every 4 days.
Zhang X., Lin H., Dong L., & Xia Q. (2022). Antiviral and Neuroprotective Abilities of Influenza Virus Infection in Tractable Brain Organoids.
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